Optics and Lasers in Engineering 50 (2012) 850–854

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Optics and Lasers in Engineering

journal homepage: www.elsevier.com/locate/optlaseng

Depolarization of light in biological tissues

Dror Fixler a,n, Rinat Ankri a, Hamootal Duadi a, Rachel Lubart b,1, Zeev Zalevsky a
a
Faculty of Engineering and the Institute of Nanotechnology and Advanced Materials, Bar Ilan University, Ramat Gan 52900, Israel
b
Department of Chemistry, Bar Ilan University, Ramat Gan 52900, Israel

a r t i c l e i n f o a b s t r a c t

Article history: Recently in phototherapy the use of diodes and broadband light devices instead of lasers was suggested
Received 1 November 2011 for economical and practical reasons. It has been argued that lasers are not superior to LEDs since they
Received in revised form lose their coherence and polarization once they penetrate into biological tissues. However, the
12 December 2011
polarization point has never been experimentally proven. In this work, to the best of our knowledge,
Accepted 13 January 2012
we have for the first time experimentally validated the conditions that affect the polarization state of
Available online 2 February 2012
light when laser illumination propagates through a biological tissue with and without flow. In our
Keywords: experiments we measured the polarization of light passing through phantoms as well as through
Flow uncooked turkey meat. The measurements were performed for varied integration time, thickness and
Phantoms
flow rates. It was experimentally validated that the tissue thickness hardly influences the polarization
Polarization
in comparison to flow for a reduced scattering coefficient of 0.8 mm  1 while there is no flow.
Tissues
Multi-Scattering Furthermore, when the flow is perpendicular to the polarization plane its velocity highly affects the
polarization. However, when the flow is parallel to the polarization plane there is almost no change in
the propagating light’s polarization state. Thus, one outcome of this work is that since the biological
tissue is not static and contains many blood vessels and capillaries, the polarization of the laser may be
lost when light penetrates the tissue.
& 2012 Elsevier Ltd. All rights reserved.

1. Introduction such as musculoskeletal injures, arthritis, chronic wounds, etc.

Light phototherapy uses monochromatic light (lasers), quasi-
The use of non-polarized light instead of polarized lasers in monochromatic radiation (LEDs) and broadband light devices.
biomedical applications such as phototherapy was recently sug- The laser differs from other light sources in its polarization
gested for economical and practical reasons. In many, although characteristics. The growing acceptance of LEDs and broadband
not all, cases the output of a laser is polarized. This normally light devices (which are non-polarized light sources) in photo-
means a linear polarization state, where the electric field oscil- therapy puts into debate the value of polarization for enhancing
lates in a certain direction perpendicular to the propagation the benefit from the light associated biomedical treatment.
direction of the laser beam [1]. It has been argued that lasers A widely discussed issue in the laser phototherapy clinical
are not preferred, as the latter lose their polarization direction community is the question whether the coherence of laser
once they penetrate into static biological tissues, all the more radiation has additional benefits, compared to monochromatic
once they penetrate into flowing medium. light originated from a conventional light source or LED of the
Light–tissue interaction is common in clinical treatments and same wavelength and intensity that is not polarized by additional
in medical research as different light sources in the visible and the filters. Our previous research deals with the preservation of the
near infrared (NIR) spectral ranges are widely used. Especially, in coherence state while propagating through a tissue with flow [6].
the therapeutic field, such as photodynamic therapy that uses There we have found that the laser coherence is partially lost in
light to damage tumor cells while light activates a photochemical proportion to the fluid’s flow rate through the tissue.
that induces the formation of reactive oxygen species that induces In this paper we intend to investigate the preservation of the
damage, and in Low Level Laser Therapy (LLLT) that uses visible– polarization state for light propagating through a tissue with and
NIR light to bio-stimulate cells [2–5]. LLLT has been used during without flow.
the last years for the treatment of various pathological conditions, Specially designed experiments in the cellular level have
presented that He–Ne laser radiation (i.e., coherent and polarized
light at 632.8 nm) and properly filtered light (633 nm) from a
n conventional light source, with the same intensity and irradiation
Corresponding author. Tel.: þ972 3 5317598; fax: þ972 3 7384051.
E-mail address: dror.fi[email protected] (D. Fixler). time, have the same biological effect [7]. The same conclusion was
1
Tel.: þ972 3 5317797; fax: þ972 3 7384053. drawn from an experiment in which radiation from a He–Ne laser

0143-8166/$ - see front matter & 2012 Elsevier Ltd. All rights reserved.
doi:10.1016/j.optlaseng.2012.01.011
 D. Fixler et al. / Optics and Lasers in Engineering 50 (2012) 850–854 851

was used with or without an optical fiber for full depolarization of In Section 2 we describe the materials and methods of the
the beam [8]. measurement system. In Section 3 we perform the experimental
To further evaluate the importance of polarization for the measurements and present the obtained results. Section 4 con-
stimulation of cellular metabolism, Karu et al. [9] studied a possible cludes the paper.
dependence of the level of light polarization (99.4%, 60.9% and 34.2%
linearly polarized light at 637 nm) on the model of cell attachment
to a glass matrix. They concluded that elementary processes such as 2. Materials and methods
stimulation of cell attachment do not depend on the light polariza-
tion level. However, this conclusion is true for a cellular suspension In this research we built an experimental set-up for polariza-
and might differ from real bio-tissues. tion measurement of the light being transmitted through phan-
Two other in-vivo works (in animal) claimed different effects toms containing intralipid and ink components [27] as well as
of polarized and non-polarized light. Chumak et al. [10] found uncooked turkey meat. Solid phantoms with different structures
that the irradiation of rat hind limb with polychromatic polarized and optical properties (absorption coefficient range of 0.0064–
light at 400–2000 nm produced a long-term inhibition of baseline 0.03 mm  1, scattering coefficient range of 2–3 mm  1 and aniso-
afferent traffic in the saphenous nerve, while non-polarized light tropy factor of g¼0.72) were prepared in order to simulate
produced biphasic changes in baseline activity. Ribeiro et al. [11] different human tissues. The turkey meat’s reflected light inten-
reported a very interesting effect concerning light polarization. sity profile was compared and found to be similar to those of the
They investigated the healing of burns in rats by irradiating the phantoms [6]. The phantoms were made using ink 0.1%, as an
lesions with a linearly polarized He–Ne laser beam parallel to the absorbing component, intralipid 20% (IL, Lipofundin MCT/LCT 20%,
rat’s spinal column and using the same laser and dose, but B. Braun Melsungen AG, Germany), as a scattering component and
aligning the light polarization perpendicularly to the relative agar powder, in order to convert the solution into gel. The
orientation of the spinal column. They found that the skin wound phantoms were prepared in cell culture plates (90 mm) and were
repair depended on polarization orientation with respect to a cooled in vacuum conditions (to avoid bubbles). The phantoms’
reference axis of the animal’s spinal column. optical properties are presented in Table 1. The reduced scattering
At the clinical level, there are a lot of studies [12–18] presenting coefficient ms0 (ms0 ¼ ms(1  g)) that we used was relatively small
successful polarized light therapy at 400–2000 nm, 40 mW/cm2, (0.8 mm  1) in order to preserve the polarization of the propagat-
2.4 J/cm2, for wound healing, pain relief and more. Unfortunately, ing light in the static measurements. Typically, one chooses a
almost all of these studies did not compare their results with the phantom by matching its scattering and absorption properties, to
same device having non-polarized radiation. Only Zhevago et al. those of the tissue. However, this does not necessarily imply that
[19] performed a randomized, placebo-controlled double-blind trial the polarization properties of the phantom match those of the
study with the same polarized and non-polarized light source. They tissue [28], which is the goal in our experiments. Note that a
measured changes in the humeral immunity of a large group of recent work showed that such intralipid based phantoms, as used
volunteers after exposure of a small body area to polychromatic in our study, highly imitate depolarization tissue properties [29].
visible and infrared polarized (VIP) and non-polarized (VInP) The experimental system includes (Fig. 1a) a green doubled
light (400–3400 nm, 95% polarization, 40 mW/cm2, 12 J/cm2 and Nd:YAG laser at a wavelength of 532 nm as an excitation source
400–3400 nm, no polarization, 38 mW/cm2, 11.2 J/cm2). They that was used to illuminate the sample after passing through a
demonstrated high similarity between the effects induced by a polarizer. The light that passes through the sample is captured
single exposure of both light sources. using a Pixelink PL-A741-E camera (PixeLINK, Ottawa, ON) with a
Another type of static measurement that found a difference pixel size of 7 mm  7 mm after passing through a second polar-
between polarization directions at in-vivo measurement was izer. The distance between the excitation polarizer and the
reported 20 years ago [20]. Anderson separated light reflected from flowing medium was 6 cm and the detector was focused on the
skin into two components: regular reflectance or ‘‘glare’’ arising emission polarizer (real distance of 2.5 cm). The camera lens’
from the surface and light backscattered from within the tissue. He numerical aperture (NA) was 0.2. The optical setup diagram
found that when the planes of polarization are parallel, images with (Fig. 1b) illustrates that since the illumination is scattered by
enhanced surface details are obtained. However when the planes the sample, there is a need for a camera lens in order to collect the
are orthogonal, wrinkles and surface details disappear and an light to the CCD detector.
enhanced view of vasculature and pigmented lesions is obtained. The flows inside the phantoms were generated using needles
Although it is known that polarized light is rapidly scrambled with two different diameters of G21 and G22 (G-Gauge; resulting in
in highly scattering media such as biological tissues (probably in 0.8128 mm and 0.7112 mm, respectively). The actual diameter of the
the first few hundred mm) [21,22], it seems very unlikely that tunnel in the phantom and the tissue was verified using an optical
polarization could play a role in any significant applications for microscope (Olympus BX51) once the needles were withdrawn.
the upper skin layers. The ability of polarized light to aid tissue The tunnels inside the samples (phantoms as well as the
imaging has a long history that was summarized by Jacques et al. turkey meat) were created by needles and water was injected to
[23]. Sankaran et al. [24,25] studied the influence of densely the tunnels by a 10 ml syringe. In order to get different flow
packed microsphere solution on the transmitted polarized beam,
demonstrating that the behavior of polarized light in tissues is Table 1
similar to the behavior in densely packed microsphere solutions. Optical properties of the irradiated phantoms. The concentration of IL refers to the
A few years later they showed that circularly polarized light is fraction of solids in the solution, while the concentration of ink pertains to the
always less depolarized than linearly polarized states unless the fraction of the original product.
relative degree of polarization values of both polarizations highly
Phantom Optical properties Ink concentration IL concentration
depends on the scattering regime, which is typical for the Mie # [mm  1] [%] [%]
regime and multiple scattering [26]. However, further experi-
ments with bio-tissues are needed in order to acquire higher 1 m ¼0.0064, ms0 ¼0.8 2  10  4 0.8
understanding of the possible role of the polarization of light in 2 m ¼0.0073, ms0 ¼0.8 2.2  10  4 0.8
3 m ¼0.0096, ms0 ¼0.8 3  10  4 0.8
phototherapy. In the present study we experimentally tested the 4 m ¼0.0192, ms0 ¼0.8 6  10  4 0.8
loss of polarization in tissues with and without flow.
852 D. Fixler et al. / Optics and Lasers in Engineering 50 (2012) 850–854

particulates, the light’s polarization status changes. For this reason,

first we measured the polarization of light that passed through the
scattering and absorption medium of the static phantoms. We tested
different types and widths of phantoms. The phantoms were pre-
pared as described in Section 2 and four different phantoms that are
presented in Table 1 were measured. The samples were prepared in
order to mimic the optical properties of real biological tissues. Each
sample was measured for varied widths of 1, 2, 4 and 5 mm.
In Fig. 2 one can see representative results for the 4 different
phantoms (from Table 1). The polarizers were positioned perpen-
dicularly to one another, so that when no sample was inserted
between them no light was collected by the camera and the
polarization state (the normalized intensity along the direction of
the output polarizer) remained 1 (marked as sample #0 in Fig. 2).
When inserting the samples (1, 2, 4 and 5 mm wide phantoms
with the characteristics described in Table 1), one could see a
small reduction in the polarization state, but in all cases the
polarization remained higher than 0.92. These results are in high
agreements with previous reports [23,24].

3.2. Polarization of real tissues

Solid phantoms composed of ink and interlipid suspended in

Fig. 1. Image (a) and optical diagram (b) of the experimental setup. A laser water may have bulk optical properties similar to those of a tissue
illuminates a sample after passing through the left polarizer and the transmitted (absorption and scattering coefficients, anisotropy of scattering,
light is imaged by a camera after passing through the right polarizer. etc), but one may claim that they will not likely recapitulate
tissue properties. The reason might be that the nature of the
refractive index variation in the phantom is caused by discrete
velocities, we used 1, 1.5 and 2 ml of tap water, purified water, particulates, while tissue is a random continuum. In addition, it
PBS (pH¼7.4) and Dulbecco’s modified Eagle’s medium (including does not necessarily imply that the polarization properties of the
Calcium, Magnesium, Potassium and Sodium), which were phantom match those of the tissue [28].
injected to the tunnels by an engine bundled to different syringes, In order to test this issue, measurements of real tissues (pieces
where the content of the syringe was injected to the sample of uncooked turkey meat) were performed and compared to
during the same period of time (10 s). The scattering properties of phantom samples with similar optical properties [32]. The mea-
the tap water, purified water and PBS were zero; the optical surements were done for different widths of tissues and phan-
properties of Dulbecco’s modified Eagle’s medium were not toms (1–4 mm). As can be seen from Fig. 3, the polarization
measured directly but before starting the flow, the first control properties in these tissues are identical to those measured in
measurement was done including the sample and the medium phantoms. Since scattering in static tissue structures rapidly
without flow. We planned the above flow velocities (6–12 ml/s), depolarizes an incident beam, we have chosen to use samples
volumes and tunnel diameters in order to achieve typical human whose polarization state remained high ( 40.9) without flow and
blood flow velocities. The blood flow in the entire human only the flow depolarization takes effect.
circulation is about 5000 ml/min in an average sized adult at
rest, but may be 5–6 times greater during heavy exercise [30]. The
3.3. Flow inside the phantoms
blood vessel diameters change from a few mm down to microns,
so our rates and diameters are similar to the neck vein blood flow
Measurements of flow in phantoms were done as follows:
velocity [31].
phantoms with the same characteristics as were used for the
By the term of polarization we refer in this research to the
following ratio of measured intensities:
I0,0 I0,90
P¼
I0,0 I0,90

where Ia,b is the intensity measured when the first polarizer was
positioned at an angle a and the second at an angle b.
The captured videos of the transmitted light were stored as avi
files and analyzed using the MATLAB (The MathWorks, Natick,
MA) software. In particular, avi movies were translated to
individual frames where each one of them contains all the image
information, such as its time and exposure duration.

3. Experimental results

3.1. Static phantom

Fig. 2. Polarization measured by the experimental setup for different phantoms
without flow. Four different phantoms with 4 mm width are presented and their
As polarized light propagates through light-scattering media, such optical properties are described in Table 1. The polarization of all samples is larger
as biological tissues, microsphere solutions or an atmosphere with than 0.92.
 D. Fixler et al. / Optics and Lasers in Engineering 50 (2012) 850–854 853

Fig. 3. Polarization for phantoms (diamonds) and for a piece of turkey meat
(square) with different widths. Note that both samples have a contrast close to
one, similarly to what was obtained in Fig. 2.
Fig. 4. Polarization for phantom with different flow conditions. Three different
flow rates were measured for three different orientations of the flow-polarizer.
The full thin lines present a flow velocity of 0.1 ml/s; the dashed lines present a
static phantom measurements and with 5 mm width were used velocity of 0.15 ml/s; the full thick lines present a velocity of 0.2 ml/s; the black
for the flow measurements. Two different samples were pre- lines present flow in the same direction of the polarizer; the red lines present flow
at 451 to the polarizer; the green lines present flow perpendicular to the direction
pared: one sample with a single tunnel of 0.8128 mm in diameter
of the polarizer. (For interpretation of the references to color in this figure legend,
and another sample with a single tunnel of 0.7112 mm in the reader is referred to the web version of this article.)
diameter. The tunnels were generated in the middle of the
samples. Four types of liquids (tap water, purified water, PBS
and Dulbecco’s modified Eagle’s medium) at flow rates of 0.1 ml/s, flowing in many directions. Therefore, we measured the polariza-
0.15 ml/s and 0.2 ml/s were injected into the tunnels. The flows tion of light propagating through a biological tissue versus the
were in different directions relative to the excitation polarizer. volumetric flow rate and the integration time as well as the flow
We measured flow in the same direction as that of the polarizer direction. We found that the polarization of the laser is almost
excitation, in a perpendicular direction to the polarizer and at 451. uninfluenced by light passing through a static tissue with a low
The results for the polarization state obtained for the purified reduced scattering coefficient of ms0 ¼0.8 mm  1 or when the flow
water are presented in Fig. 4. When the polarizer and the flow were direction is parallel to the polarization direction of the laser.
in the same direction there was almost no decrease in the polariza- However, it is partially lost when there is a flow of fluid through
tion (still higher than 0.9) detected for all flow rates (black lines). the tissue in a direction perpendicular to the laser’s polarization
When the flow was in a perpendicular direction to the light, the direction. No evidence for optical activity was found. It should be
polarization state decreased up to 0.4 (green lines). For different noted that in the current study the tissues that we used was fresh
flow rates we got different integration durations that generated a flash tissues, which did not mimic the actual in vivo situation
reduction of the polarization; for a flow rate of 0.1 ml/s in the where blood is flowing within the living tissue.
perpendicular direction the polarization was reduced after 2 s to 0.6 The volumetric flow rate is directly correlated to the polariza-
(full thin green line), while for 0.2 ml/s flow velocity in the tion loss. Higher flow rate produces lower polarization values.
perpendicular direction the polarization reduced after 2 s to its final This may happen due to the flow itself as well as due to the
value of 0.4 (full thick green line). When the flow was at 451, we got movements of the scattering elements inside the tissues, caused
the polarization values between the last two cases (red lines). For by the flow. The multi-scattering phenomena reduced the polar-
the other three types of liquids (tap water, PBS and Dulbecco’s ization value. The fact that the blood vessels in the body do not
modified Eagle’s medium) the results were the same (data not range in one direction means that the laser beam necessarily hits
shown). the fluid flow through the tissue perpendicular to its polarization
These results, dealing with the different orientation of the direction.
polarizer relative to the flow direction, match a mathematical relation It can thus be concluded that from the polarization aspect –
of cos2 y between the intensity in the perpendicular direction and the based on the fact that linear polarization of laser radiation after
relative angle, meaning that the flow will not influence the polariza- passing fluid media may be lost, there are no additional benefits
tion when it is in the same direction as the incident light’s polariza- in using it as an excitation source without maintaining the
tion. However, when there is an angle of 901 between the polarization excitation direction; for example it may simplify pulse oximetry
and the flow direction, the depolarization effect will be significantly instrumentation. Furthermore, the change in polarization as a
stronger and at 451 the polarization is reduced by half. While at 01 we flow rate indicator could be used as a diagnostic tool for
measure P¼ 0.9, at 901 P¼0.4; hence DP¼P(y ¼01)—P(y ¼901)¼0.5 congestive heart failure. It might also follow wound healing rates
and P(y ¼ 451)¼0.65 is exactly DP cos2 451þ P(y ¼901). in diabetic patients.

4. Conclusions and discussions

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